Alterations in salivary biochemical composition and redox state disruption induced by the anticonvulsant valproic acid in male rat salivary glands.

OBJECTIVE: To investigate the effects of the anticonvulsant valproic acid (VPA) on salivary glands in male rat using biochemical, functional, histomorphometric, and redox state parameters. MATERIALS AND METHODS: Twenty-four male Wistar rats were randomly distributed into three groups (n = 8 per group): Control (0.9% saline solution), VPA100 (100 mg/kg), and VPA400 (400 mg/kg). After 21 consecutive days of treatment with by intragastric gavage. Pilocarpine-induced saliva was collected to determine salivary flow rate, pH, buffering capacity, and biochemical composition. Analyses of histomorphometric parameters and redox balance markers were performed on the parotid and submandibular glands. RESULTS: Salivary flow rate, pH, buffering capacity, total protein, potassium, sodium, and chloride were similar between groups. However, phosphate and calcium were reduced in VPA400, while amylase was increased in both VPA100 and VPA400. We did not detect significant differences in the areas of acini, ducts, and connective tissue in the salivary glands between the groups. There were no significant changes in the redox status of the submandibular glands. In turn, in the parotid glands we detected reduced total oxidizing capacity and lipid peroxidation, measured as thiobarbituric acid reactive substances (TBARs) and higher uric acid concentration in both the VPA100 and VPA400 groups, and increased superoxide dismutase (SOD) in the VPA400 group. CONCLUSION: Chronic treatment with VPA modified the salivary biochemical composition and caused disruption in the redox state of the parotid gland in rats.

Assessment of the toxic effects of levetiracetam on biochemical, functional, and redox parameters of salivary glands in male Wistar rats.

Levetiracetam (LEV) is an anticonvulsant for epilepsy. The toxic effects of this medication in tissues have been associated with redox state imbalance, which can lead to salivary gland dysfunction. Therefore, the current work investigated the effects of LEV on the biochemical, functional, and redox parameters of the parotid and submandibular glands in rats. For this, male Wistar rats (Rattus norvegicus albinus) were randomly divided into 3 groups (n = 10/group): Control (0.9% saline solution), LEV100 (100 mg/kg), and LEV300 (300 mg/kg). After 21 consecutive days of intragastric gavage treatments, pilocarpine stimulated saliva secretion was collected for salivary biochemical analysis. The extracted salivary glands were utilized for histomorphometry and redox state analyses. Our results showed that LEV300 increased plasma hepatotoxicity markers and reduced salivary amylase activity and the acinar surface area of the parotid gland. Total oxidant capacity and oxidative damage to lipids and proteins were higher in the parotid gland, while total antioxidant capacity and uric acid levels were reduced in the submandibular gland of the LEV100 group compared to Control. On the other hand, total oxidant capacity, oxidative damage to lipids and proteins, total antioxidant capacity, and uric acid levels were lower in both salivary glands of the LEV300 group compared to Control. Superoxide dismutase and glutathione peroxidase activities were lower in the salivary glands of treated animals compared to Control. In conclusion our data suggest that treatment with LEV represents a potentially toxic agent, that contributes to drug-induced salivary gland dysfunction.

Assessment of redox state and biochemical parameters of salivary glands in rats treated with anti-obesity drug sibutramine hydrochloride.

OBJECTIVES: To investigate the effects of anti-obesity drug sibutramine hydrochloride (SB) on redox state and biochemical parameters in the salivary glands. MATERIALS AND METHODS: Adult male Wistar rats were randomly divided into the following groups (n = 8 per group): control rats treated with vehicle (C) and rats treated with SB (10 mg/kg/day) by intragastric gavage for 28 days. The parotid (PG) and submandibular (SMG) glands were processed using histomorphometric analysis, and total protein, amylase, mucin, and oxidative damage to lipids were determined by measuring the formation of thiobarbituric acid reactive substances (TBARS), total antioxidant capacity (TAC), uric acid (UA), total glutathione (tGSH), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), and AKT phosphorylation. RESULTS: SB decreased the acinar area, and increased the stromal area in PG, while no effect on the morphometric parameters was observed in SMG. SB also increased oxidative damage to lipids (TBARs). The SB group showed lower total protein, amylase, TAC, UA, tGSH, SOD, CAT, and GPx than the C group in PG, while in SMG, SB decreased total protein, mucin, tGSH, SOD, CAT, and GPx. However, increased AKT phosphorylation observed in both salivary glands suggests that SB exerts low-intensity oxidative stress. CONCLUSIONS: SB impaired enzymatic and non-enzymatic antioxidant defenses in the salivary glands of rats. CLINICAL RELEVANCE: Chronic treatment with SB could mitigate salivary gland dysfunction due to disturbance of redox state.

Effects of orchiectomy and testosterone replacement therapy on redox balance and salivary gland function in Wistar rats.

The objective of this study was to investigate the effects of orchiectomy (ORX) and testosterone replacement therapy (TRT) on redox balance and function of salivary glands. Forty-five young adult male Wistar rats (3 months old) were either castrated bilaterally or underwent fictitious surgery (SHAM) and were subsequently distributed into 3 groups: SHAM, ORX, and TRT (castrated rats that received an intramuscular injection of testosterone cypionate 10 mg/kg/weekly). All treatments started 4 weeks after castration (4 months old) and lasted 4 weeks (5 months old). At the end of treatment, pilocarpine-induced salivary secretion was collected to analyze salivary flow rate and biochemistry composition, and the parotid (PG) and submandibular (SMG) glands were sampled for redox balance markers and histomorphometric analyses. ORX increased salivary flow rate, calcium, phosphate, and chloride, and decreased total protein and amylase, while not changing the salivary buffer capacity, pH, sodium, and potassium compared to SHAM. TRT restored all salivary parameters to SHAM values. ORX increased oxidative lipid and protein damage, total antioxidant capacity, and uric acid in both salivary glands compared to SHAM. Superoxide dismutase, catalase, and glutathione peroxidase activities were greater only in the SMG of the ORX group in relation to SHAM. ORX decreased duct and acini area, while increasing connective tissue in the PG. On the other hand, ORX reduced duct area and increased acini area in the SMG compared to SHAM. TRT restored the redox balance and histomorphometric parameters to close to SHAM values in both salivary glands. Orchiectomy-induced salivary gland dysfunction was characterized by an increase in the salivary flow rate and changes in the secretion of total protein, amylase, and electrolytes, which are key factors, considered important for maintaining oral health status. To sum up, orchiectomy impaired the redox balance of the salivary glands. Our results also showed that TRT reversed the oxidative damage, morphological alterations, and salivary gland dysfunction induced by orchiectomy. Therefore, these results suggest an important action of testosterone on the redox balance and secretory ability of salivary glands.

Salivary flow and salivary proteins in male and female children with autism spectrum disorder: pilot study / Fluxo salivar e proteínas salivares em crianças dos gêneros masculino e feminino com transtorno do espectro do autismo: estudo piloto

Introduction: autism spectrum disorder is a neurodevelopmental condition that affects the establishment of bonds and communication. Dental care is more difficult for people with this disorder, because in addition to communication difficulties, non-cooperation with respect to oral hygiene and continuous use of medication are common. Greater predisposition to caries, as well as alterations in the flow and concentration of salivary proteins were reported in these individuals. Objective: considering that sex can affect salivary flow and protein concentration, our objective was to analyze these parameters in the saliva of children with autism. Material and method: total unstimulated saliva was obtained from 12 boys and 12 girls aged between 5 and 15 years, with the aid of a catheter, after 2 hours of fasting and oral hygiene. Salivary flow was determined by estimating the mass of saliva. Total protein was determined in the supernatant obtained after centrifugation at 10,000 x g, for 10 minutes, by the Lowry method, with bovine albumin as standard. The results are expressed as mean and standard deviation. The data were submitted to the Shapiro-Wilk and Mann Whitney tests, with a significance level of 5%. Result: salivary flow values for boys (0.3555 ± 0.24 ml/min) and girls (0.2522 ± 0.1727 ml/min), and protein values for boys (1.430 ± 0.7480 mg/mL) and girls (1.075 ± 0.3702 mg/mL) were not significantly different between sexes. Conclusion: in children with autism spectrum disorder, sex does not influence unstimulated flow and salivary protein values.

Introdução: o transtorno do espectro autista é uma condição do neurodesenvolvimento que afeta o estabelecimento de vínculos e a comunicação. Cuidados odontológicos são mais difíceis em portadores desse transtorno, pois além da dificuldade de comunicação são comuns a não cooperação na higiene bucal e uso contínuo de medicamentos. Maior predisposição à cárie, alterações no fluxo e na concentração de proteínas salivares foram relatadas nesses indivíduos. Objetivo: considerando que o gênero pode afetar fluxo salivar e concentração proteica, nosso objetivo foi analisar esses parâmetros na saliva de crianças com autismo. Material e método: saliva total não estimulada foi obtida de 12 meninos e 12 meninas com idades entre 5 e 15 anos, com auxílio de um cateter, após 2 horas de jejum e higienização da cavidade bucal. O fluxo salivar foi determinado estimando-se a massa de saliva e o total de proteínas foi determinado no sobrenadante obtido após centrifugação a 10.000 x g, por 10 minutos, pelo método de Lowry, com albumina bovina como padrão. Os resultados foram expressos como média e desvio padrão, sendo submetidos aos testes de Shapiro-Wilk e Mann Whitney, com nível de significância em 5%. Resultado: os valores de fluxo salivar não foram significativamente diferentes quando comparados meninos (0.3555 ± 0.24 ml/min) e meninas (0.2522 ± 0.1727 mL/min), bem como os valores de proteínas (meninos: 1.430 ± 0.7480 mg/mL; meninas 1,075 ± 0,3702 mg/mL). Conclusão: em crianças com transtorno do espectro autista o gênero não influencia os valores de fluxo não estimulado e proteínas salivares.

Análise bioquímica da saliva de pacientes com deficiência cognitiva e física como estratégia de aula prática de Bioquímica no curso de Odontologia / Biochemical analysis of the saliva of patients with cognitive and physical disabilities as a practical Biochemistry class strategy in the Dentistry course

Diante da importância do ensino de Bioquímica na formação do futuro cirurgião-dentista, o objetivo é apresentar uma estratégia pedagógica que possibilita a articulação dos conceitos moleculares básicos da disciplina no entendimento da complexidade das doenças bucais e sistêmicas, por meio da análise bioquímica da saliva de pacientes com deficiência cognitiva e física do Centro de Assistência Odontológica à Pessoa com Deficiência (CAOE) da Faculdade de Odontologia de Araçatuba-Universidade Estadual Paulista(UNESP).Para tanto, grupos de cinco estudantesrealizaram as análises na saliva, coletada por um cirurgião-dentista doCAOE. Imediatamente após a coleta, foi determinado o fluxo salivar, pH e a capacidade tamponante. Na sequência, a saliva foi centrifugada, fracionada e armazenada a -20 °C até o momento das análises bioquímicas. Durante as aulas práticas, realizaram-se os seguintes ensaios, utilizando kitscomerciais: proteína total, α-amilase, fosfatase alcalina, ácido úrico, glicose, aspartato aminotransferase, cálcio e fósforo. Ao final do ano letivo, os estudantes apresentaram relatório contextualizando os resultados com a saúde bucal e sistêmica do paciente. Plantões de dúvidas com monitores e professores auxiliaram na interpretação da ficha de anamnese e nas correlações dos parâmetros clínicosbucais e sistêmicos. Para os estudantes ingressantes, foi a primeira oportunidade de integrar o conhecimento teórico às condições clínicas de um paciente. Conclui-se que a estratégia adotada é viável e pode beneficiar educadores que buscam alternativas que permitam a integração das ciências básicas e clínicas ao ensino de Bioquímica para a Odontologia (AU).

Given the importance of teaching biochemistry in dental surgeon trainingcourses, the objective was to present a pedagogical strategy that enables the articulation of basic molecular concepts within the subject, allowing a betterunderstanding of the complexity of oral and systemic diseases, through the biochemical analysis of saliva from patients with cognitive and physical disabilities at the Dental Assistance Center for Disabled Persons (CAOE)of the School of Dentistry, Araçatuba -São Paulo State University (UNESP).For this purpose, groups of five students analyzedthesaliva collected by a dentalsurgeon from CAOE. Immediately after collection, salivary flow, pH, and buffering capacity were determined. Subsequently, the saliva was centrifuged, fractionated, and stored at -20°C until the time of biochemical analysis. During the practical classes, using commercial kits, the samples were tested for: total protein, α-amylase, alkaline phosphatase, uric acid, glucose, aspartate aminotransferase, calcium, and phosphorus. At the end of the school year, students presented a report contextualizing the results with the patient's oral and systemic health. Tutoring sessions with monitors and teachers helped in the interpretation of the anamnesis form and the correlations of oral and systemic clinical parameters. For incoming students, this was the first opportunity to integrate theoretical knowledge with clinical perspectives.It is concluded that the adopted strategy is viable and can benefit educators who seek alternatives that allow the integration of basic and clinical sciences forteaching Biochemistry in Dentistry (AU).

The effects of storage time and temperature on the stability of salivary phosphatases, transaminases and dehydrogenase.

OBJECTIVES: To investigate the influence of temperature and storage time on salivary acid phosphatase (ACP), tartrate-resistant acid phosphatase (TRAP), alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and lactate dehydrogenase (LDH). DESIGN: Unstimulated whole expectorated saliva was collected from healthy men and women subjects (n=26) between 8 and 10a.m. The saliva samples were centrifuged, and the supernatants were measured for ACP, TRAP, ALP, AST, ALT and LDH activities immediately (without freezing) [baseline values] and after time intervals of 3, 7, 14 and 28days (d) of storage at -20°C and -80°C using spectrophotometric methods The influence of storage time was analyzed by one-way ANOVA followed by the Dunnett post-test, while the paired Student's-t-test was used to compare the differences between the temperature (p<0.05). RESULTS: There was significant decline in the activities of all enzymes at -20°C with increasing storage time. This decrease was relevant from day 14 onward for the majority of the enzymes, with the exception of AST. After day 28, the more sensitive enzymes were ALP and LDH, which showed residual activity of 39% and 16%, respectively, compared with baseline values. There were considerable, but insignificant changes, in the activities of all enzymes after storage at -80°C for 28days. CONCLUSIONS: Frozen samples should be kept at -80°C to preserve these activities, but there are restrictions for the enzymes ALP, ALT and LDH. Storage of samples at -20°C could introduce high error variance in measured activities.

Yacon (Smallanthus sonchifolius) Leaf Extract Attenuates Hyperglycemia and Skeletal Muscle Oxidative Stress and Inflammation in Diabetic Rats.

The effects of hydroethanolic extract of Yacon leaves (HEYL) on antioxidant, glycemic, and inflammatory biomarkers were tested in diabetic rats. Outcome parameters included glucose, insulin, interleukin-6 (IL-6), and hydrophilic antioxidant capacity (HAC) in serum and IL-6, HAC, malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) in soleus. The rats (10/group) were divided as follows: C, controls; C + Y, HEYL treated; DM, diabetic controls; and DM + Y, diabetic rats treated with HEYL. Diabetes mellitus was induced by administration of streptozotocin. C + Y and DM + Y groups received 100 mg/kg HEYL daily via gavage for 30 d. Hyperglycemia was improved in the DM + Y versus DM group. Insulin was reduced in DM versus C group. DM rats had higher IL-6 and MDA and lower HAC in the soleus muscle. HEYL treatment decreased IL-6 and MDA and increased HAC in DM rats. DM + Y rats had the highest CAT activity versus the other groups; GPx was higher in C + Y and DM + Y versus their respective controls. The apparent benefit of HEYL may be mediated via improving glucoregulation and ameliorating oxidative stress and inflammation, particularly in diabetic rats.

llex paraguariensis decreases oxidative stress in bone and mitigates the damage in rats during perimenopause.

During perimenopause, oxidative stress increases, which may result in disruption of bone turnover, and consequently in osteoporosis. The use of antioxidants may be an effective nutritional approach to reducing osteoporosis in this period of life. Mate tea (MT) (Ilex paraguariensis), a typical and inexpensive beverage consumed in the Brazilian south-east, Argentina and Uruguay, increases antioxidant defense. Our hypothesis was that MT would decrease oxidative stress and mitigate bone deterioration. To test this, we analyzed oxidative stress markers of bone turnover, and local and systemic markers of bone metabolism of rats during natural perimenopause. Female Wistar rats (aged 16months) in proven perimenopause period received 20mg/kgBW/day of mate tea, by gavage (PM+MT Group, n=10) or water (PM Group, n=10). Female rats aged 4months (AD Group, n=10) received water. The treatment period was four weeks. MT minimized the deterioration of rat microarchitecture, characterized by increase in the bone trabecular area, number of osteocytes and areal bone mineral density. These results were accompanied by a lower level of malondialdehyde, an oxidative stress marker, in femoral tissue homogenate. Plasmatic tartrate-resistant acid phosphatase, a typical osteoclastic function marker, decreases after treatment, indicating a decrease in osteoclastic function. MT also modified the immunostaining pattern of bone metabolism markers, decreasing the receptor activator of nuclear factor kappa-B ligant (RANKL), superoxide dismutase isoform 2 (SOD2) and increasing osteoprotegerin (OPG), a decoy receptor for the RANKL, which positively modulates bone mass. These results suggested MT was capable of decreasing bone resorption by inhibiting the osteoclastogenesis in a RANKL-dependent signaling pathway activated by oxidative stress. Taken together, the results indicated that MT minimized bone loss in perimenopause and this effect is at least partly due to the decrease in oxidative stress, confirming our hypothesis.

Ilex paraguariensis supplementation may be an effective nutritional approach to modulate oxidative stress during perimenopause.

Perimenopause is a period in a woman's life that precedes menopause and is characterized by hormonal changes that result in increased oxidative stress. Since oxidative stress is associated with age-related diseases and perimenopausal symptoms including somato-vegetative manifestations, nutritional antioxidant supplementation may be an effective approach to minimizing this stress. Mate tea (MT) (Ilex paraguariensis), a typical and inexpensive beverage consumed in the Brazilian south-east, Argentina and Uruguay, increases antioxidant defense. We hypothesized that MT could minimize oxidative stress during perimenopause by modulating enzymatic antioxidant defense. To test this, we analyzed the lipid oxidative damage and antioxidant defense in erythrocytes and liver of rats, after MT treatment. Female Wistar rats (aged 16months) in proven perimenopause period received 20mg/kgBW/day of mate tea, by gavage (PM+MT group) or water (PM group). Female rats aged 4months (AD group) received water. Erythrocytes and liver were used to determine lipid oxidative damage, determined by malondialdehyde (MDA); superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT) activities. Total plasma antioxidant capacity was examined by ferric reducing antioxidant power assay (FRAP) and estrogen by radioimmunoassay. MT increased FRAP and did not change estrogen levels. Increased SOD and GPx, and reduced MDA were observed in both tissues studied. Increased CAT activity was observed only in the liver. We confirmed the hypothesis that MT was capable of minimizing oxidative stress in this period of life by modulating antioxidant defense.

Enteral nutrition feeding alters antioxidant activity in unstimulated whole saliva composition of patients with neurological disorders.

Patients with neurological disorders have an increased risk of oral and systemic diseases due to compromised oral hygiene. If patients lose the ability to swallow and chew food as a result of their disorder, enteral nutrition is often utilized. However, this type of feeding may modify salivary antioxidant defenses, resulting in increased oxidative damage and the emergence of various diseases. The aim of this study was to evaluate the effects of enteral nutrition on biochemical parameters in the unstimulated whole saliva composition of patients with neurological disorders. For this, enzymatic (superoxide dismutase - SOD; glutathione peroxidase - GPx) and non-enzymatic (uric acid; ferric ion reducing antioxidant power - FRAP) antioxidant activity, as well as a marker for oxidative damage (thiobarbituric acid reactive substances - TBARS) were analyzed. Unstimulated whole saliva was collected from 12 patients with neurological disorders and tube-feeding (tube-fed group - TFG), 15 patients with neurological disorders and normal feeding via the mouth (non-tube-fed group - NTFG), and 12 volunteers without neurological disorders (control group - CG). The daily oral hygiene procedures of TFG and NTFG patients were similar and dental care was provided monthly by the same institution's dentist. All patients exhibited adequate oral health conditions. The salivary levels of FRAP, uric acid, SOD, GPx, TBARS, and total protein were compared between studied groups. FRAP was increased (p<0.05) in the NTFG (4,651 ± 192.5 mmol/mL) and the TFG (4,743 ± 116.7 mmol/mL) when compared with the CG (1,844 ± 343.8 mmol/mL). GPx values were lower (p<0.05) in the NTGF (8.24 ± 1.09 mmol/min/mg) and the TFG (8.37 ± 1.60 mmol/min/mg) than in the CG (15.30 ± 2.61 mmol/min/mg). Uric acid in the TFG (1.57 ± 0.23 mg/dL) was significantly lower than in the NTFG (2.34 ± 0.20mg/dL) and the CG (3.49 ± 0.21 mg/dL). Protein was significantly lower in the TFG (5.35 ± 0.27 g/dL) than in the NTFG (7.22 ± 0.57 g/dL) and the CG (7.86 ± 0.54 g/dL). There was no difference in the salivary flow rate and SOD between groups. Enteral nutrition in patients with neurological disorders was associated with lower oxidative damage, resulting in increased salivary antioxidant capacity. These results emphasize the importance of oral care for this population to prevent oral and systemic diseases.

Salivary alpha amylase and cortisol levels in children with global developmental delay and their relation with the expectation of dental care and behavior during the intervention.

The purpose of this study was to analyze the alpha-amylase (sAA) and cortisol levels in children with Global developmental delay (GDD) before and after dental treatment and its association with the children's behavior during treatment. The morning salivary cortisol levels and activity of sAA of 33 children with GDD were evaluated before and after dental treatment and were compared to 19 healthy children. The behavior of children with GDD during dental care was assessed by the Frankl scale. Children with GDD showed lower levels of sAA activity than healthy children, but this result was not significant. The salivary cortisol levels were similar between GDD and healthy children. GDD children showed increased levels of sAA (but not cortisol) prior to the dental treatment as compared to the post-treatment phase. GDD children who showed less favorable behavior during dental care had higher levels of sAA and salivary cortisol than GDD children with more favorable behavior, but only the sAA results were significant. In conclusion, GDD children show hyperactivity of the SNS-axis in anticipation of dental treatment which indicates the need for strategies to reduce their anxiety levels before and during dental care.

Neuroendocrine alterations impair enamel mineralization, tooth eruption and saliva in rats

A administraçäo neonatal de glutamato monossódico (MSG) em ratos provoca distúrbios neuroendócrinos que acarretam alterações em vários sistemas orgânicos. Neste trabalho avaliamos as repercussöes desse tratamento sobre dentes e glândulas salivares. Ratos machos e fêmeas recém-nascidos foram injetados com MSG (4 mgg peso corporal, s.c.) uma vez ao dia no 2, 4, 6, 8 e 10 dia após o nascimento; o grupo controle recebeu soluçäo salina no mesmo esquema. O momento da erupçäo do incisivo inferior foi determinado entre o 4 e o 10 dia de vida, e o ritmo de erupçäo foi medido entre o 43 e o 67 dia. O fluxo de saliva e o conteúdo salivar de proteína e amilase foram determinados sob estimulaçäo com pilocarpina aos 3 meses de idade. Os animais tratados com MSG mostraram reduções significativas do fluxo salivar (machos: -27 por cento; fêmeas: -40 por cento) e do peso das glândulas submandibulares (cerca de 12 por cento). Apenas em machos houve discreta reduçäo do peso corporal (7 por cento). A saliva dos animais tratados com MSG apresentou aumento da concentraçäo de proteínas totais e da atividade amilásica. A erupçäo dos incisivos inferiores ocorreu mais precocemente nos ratos tratados do que nos controles, porém a taxa de erupçäo apresentou-se significativamente reduzida. A microdureza também foi menor nos animais tratados. Nossos resultados mostram que o tratamento de ratos recém-nascidos com MSG causa um quadro definido de alterações buco-dentais no animal adulto, traduzidas por reduçäo do fluxo de saliva (sem reduçäo da síntese protéica) e distúrbios da mineralizaçäo e erupçäo dentárias. Estes dados apontam para o importante papel modulador que certos núcleos hipotalâmicos sensíveis ao MSG exercem sobre os fatores que regulam a suscetibilidade à cárie